Digest Journal of Nanomaterials and Biostructures Vol. 7, No. 2, April - June 2012, p. 469 - 475 IMPACT OF SOLUTE MOLECULAR PROPERTIES ON THE ORGANIZATION OF NEARBY WATER: A CELLULAR AUTOMATA MODEL C. MIHAIa, A. VELEAb,e, N. ROMANc, L. TUGULEAa, N. I. MOLDOVANd** aDepartment of Electricity, Solid State and Biophysics, Faculty of Physics, University of Bucharest, Romania bNational Institute of Materials Physics, Bucharest - Magurele, Romania cDepartment of Computer Science, The Ohio State University - Lima, Ohio, USA dDepartment of Internal Medicine, Davis Heart and Lung Research Institute, The Ohio State University - Columbus, Ohio, USA e"Horia Hulubei” Foundation, Bucharest-Magurele, Romania The goal of this study was the creation of a model to understand how solute properties influence the structure of nearby water. To this end, we used a two-dimensional cellular automaton model of aqueous solutions. The probabilities of translocation of water and solute molecules to occupy nearby sites, and their momentary distributions (including that of vacancies), are considered indicative of solute molecular mechanics and hydrophatic character, and are reflected in water molecules packing, i.e. ‘organization’. We found that in the presence of hydrophilic solutes the fraction of water molecules with fewer neighbors was dominant, and inverse-proportionally dependent on their relative concentration. Hydrophobic molecules induced water organization, but this effect was countered by their own flexibility. These results show the emergence of cooperative effects in the manner the molecular milieu affects local organization of water, and suggests a mechanism through which molecular mechanics and crowding add a defining contribution to the way the solute impacts on nearby water. (Received March 19, 2012; Accepted March 24, 2012) Keywords: Water, Cellular automata, Complexity, Clustering, Hyper-mobile, Hydrophobic, Biomechanical, Crowding 1. Background Individual and collective mobility of water molecules are instrumental for many chemical and biological phenomena. For example, water splitting in photosynthesis critically depends on its accessibility to the reaction center II, which it is embedded in the lipid phase of thylakoid membranes [1]. Plasma membrane, on the other side, manifests selective filtration properties for water, displaying both controlled channel-mediated transport (through aquaporins) and a direct trans-membrane flux, larger than expected from the hydrophobic effect [2]. These and many other phenomena make the understanding of cooperative effects in water solutions an enduring but unmet goal of physical (bio)chemistry. Solutions are complex systems of molecules, studied most often from the point of view of the solute. However, although the solvent is seemingly simpler in molecular structure and properties, its role in defining the properties of these systems is no less important, as increasingly shown for water, the universal biological solvent. In this regard, the notion of 'slaving' is notorious, describing the driving role of density fluctuations in nearby bulk water for controlling the large- scale stochastic movements of dissolved biomolecules [3]. This one-directional view of solvent influencing the solute was aptly criticized, even in the case of slaving [4], because there is a strong *Corresponding author: [email protected] 470 reciprocal influence between solute and solvent. For example, the molecules often have 'kosmotropic' (water organizing) or 'chaotropic' (water disorganizing) effects on water structure [5]. An example is urea, the molecular 'denaturant' used to alter solubility of proteins, yet acting on the solvent. Therefore, solutes may be found to influence in subtle ways the cooperative nature of inter-molecular interactions in the solvent. The properties of dissolved molecules could thus inversely influence those of water producing complex, non-linear causal loops. Molecular dynamic simulations with atomic resolution remain powerful tools for the study of such reciprocal effects of solutes and water. However, due to the immense computational demand (for programming the exquisitely heterogeneous biological systems, and at the stage of execution), their applicability to charting the laws that govern the bio-molecular solutions remains limited [6]. In search for a more efficient, but still reliable computational method to address these questions, we chose cellular automata. These are well known for their ability to capture the essence of emergent phenomena in complex stochastic and/or chaotic systems, while their results are comparable with those obtained by Monte Carlo simulations (see for example [7]). Two dimensional models of water are useful [8-10], although their degree of sophistication depends on the study goal. One of the simplest but powerful versions is the cellular automata model of aqueous solutions developed by Kier et al. [11-15]. This model was largely validated against the main physico-chemical properties of water and of simple aqueous solutions, including the ability to reproduce the hydrophobic effect [11,12,15]. Kier et al. verified that the molar fraction (f) of water molecules, hydrogen-bonded from none to four other molecules, satisfies the known distribution in the actual fluid [13]. In other two-dimensional models of water, up to 6 neighbors have been considered [9,10]. For our work, we implemented a version of Kier et al. [13], with modified neighborhoods. It should be stressed that this modeling does not seek a realistic representation of water or of biomolecules (better approached by molecular dynamics and other forms of simulations). This model is designed to capture instead the general phenomena emerging in complex systems, with a new focus on molecular biomechanics, an aspect of biological systems largely neglected so far in favor of their biochemical properties. Using this model, we explored the impact of three essential properties of the biomolecules on the structure of bulk nearby water: a) the concentration (equivalent of the ‘crowding’ of cytoplasm), b) the hydrophatic character (i.e hydrophobic vs. hydrophilic) of solute molecules, and c) the molecular flexibility (ad-hoc defined in this study, see below). Our data show a surprisingly strong impact of solute concentration and of its ‘stiffness’ on the packing of nearby water. These results recapitulate the main known features of water solutions studied with more refined methods, but in a more general and more comprehensive way. This effort lays out the foundation for application of our modeling in subsequent studies to other complex phenomena in aqueous biomolecular solutions (such as ‘slaving’ and stochastic molecular mechanics). 2. Model and methods The model used throughout this study is an extension of the work of Kier et al. [11-15]. It is a two - dimensional cellular automaton, where the computing objects (or particles, representing water and solute) occupy discrete states on the grid sites. The squared sites of the grid are well suited for the investigation of water-related phenomena, because water molecules can participate in up to four hydrogen bonds with the neighbors. Each object interacts only with its immediate neighbor, so the rules that govern their behavior are local. There are several modes to consider an object neighborhood used in cellular automata (Fig. 1). The most common type is the so called ‘von Neumann neighborhood’. This neighborhood of a site (i) is composed of the four j sites placed crosswise. The ‘extended von Neumann neighborhood’ is represented by the second-order k sites, with the same type of placement. Another version, used in this model, is the ‘Moore neighborhood’ (with eight places), that includes the first-order sites m found in the corners of the i-centered site. Here we computed the fractions of molecules with none to eight neighbors in the final state, as derived from a Moore-type neighborhood (unlike Kier et al. model, where all molecules 471 were treated in von Neumann variant [15]). This was preferred because water molecules may crowd closer to each other than in ice (the ideal distribution along the tetrahedral orbitals of oxygen, projected onto the von Neumann lattice), to form, at room temperature, clusters with up to 8 water molecules [16]. Moreover, there are situations where water is biochemically accepted for processing in even larger clusters by enzymes [17], or by the photosystem II [1]. In addition, this approach allows a more uniform treatment of both water and solutes with larger degrees of engagement with the nearby molecules (for many biomolecules, obviously higher than four neighbors). k m j m k j i j k m j m k Fig. 1: Types of neighborhoods of site i, used in cellular automata [11]. Following [15], we consider two main parameters for this model: a) The breaking probability PB(XY) is the probability of a particle, X, in site i, to break away from a particle Y, in a site j, when there is exactly one occupied j site. PB(XY) can take values between 0 and 1. Low values of PB(XY) indicate a strong cohesion between X and Y, while high values reflect little interaction. b) The joining parameter J(XY) is the propensity of particle X in site i, to move toward the particle Y in site k, when the intermediate site j is vacant. This parameter can take positive real values. When J(XY) = 1, the particle is not influenced by its second order neighbors. The case J(XY) > 1 simulates a short-range (i.e. involving only the adjacent sites) attraction. Situation J(XY) < 1 can be considered as a mutual repulsion. When J(XY) = 0, the particle in site i, cannot move toward the particle in site k. The studies of Kier et al.[11-15] have shown that for the model to behave appropriately, PB should be set to 0.2 for pure water. Importantly, it was established that J can be related to PB in the case of water, through the following relationship (and thus we can use one rule for particle movement to model the system, the other being automatically derived from it): logJP 1.5B 0.6 (1) Specifically, for water where PB(WW) = 0.2 this means J(WW) = 2.